Abstract
Kwakhurin (Kwa) is a plant secondary metabolite solely present in Pueraria candollei var. mirifica (P. candollei), which has long been used as a Thai traditional herb for estrogen replacement therapy. Recently, health hazards have arisen in Japan regarding P. candollei-derived products containing potent estrogenic compounds. Therefore, the development of standardization methods for P. candollei materials is an urgent problem requiring resolution. The enzyme-linked immunosorbent assay (ELISA) is an effective analytical technique because it enables the development of sensitive and specific assays of the target compound through antigen−antibody reaction. Here, we produced a monoclonal antibody against Kwa (MAb 11F) by immunizing Kwa-bovine serum albumin (BSA) conjugates prepared using an N,N′-carbonyldiimidazole (CDI) mediated method. Stability and cross-reactivity tests of MAb 11F revealed that the MAb 11F is stable for at least 4 months at 4 °C and is highly specific to Kwa. The detectable concentration range of an indirect competitive ELISA (icELISA) using MAb 11F exhibited values of 1.53–48.8 ng/mL with the limit of detection (LOD) of 1.13 ng/mL. Validation analyses revealed that the developed icELISA is precise, accurate, and reliable enough to be applied to P. candollei-derived samples and products for their standardization.
| Original language | English |
|---|---|
| Pages (from-to) | 23-28 |
| Number of pages | 6 |
| Journal | Fitoterapia |
| Volume | 133 |
| DOIs | |
| Publication status | Published - Mar 1 2019 |
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All Science Journal Classification (ASJC) codes
- Pharmacology
- Drug Discovery
Cite this
An indirect competitive enzyme-linked immunosorbent assay toward the standardization of Pueraria candollei based on its unique isoflavonoid, kwakhurin. / Yusakul, Gorawit; Togita, Rui; Minami, Kei; Chanpokapaiboon, Kaskamol; Juengwatanatrakul, Thaweesak; Putalun, Waraporn; Tanaka, Hiroyuki; Sakamoto, Seiichi; Morimoto, Satoshi.
In: Fitoterapia, Vol. 133, 01.03.2019, p. 23-28.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - An indirect competitive enzyme-linked immunosorbent assay toward the standardization of Pueraria candollei based on its unique isoflavonoid, kwakhurin
AU - Yusakul, Gorawit
AU - Togita, Rui
AU - Minami, Kei
AU - Chanpokapaiboon, Kaskamol
AU - Juengwatanatrakul, Thaweesak
AU - Putalun, Waraporn
AU - Tanaka, Hiroyuki
AU - Sakamoto, Seiichi
AU - Morimoto, Satoshi
PY - 2019/3/1
Y1 - 2019/3/1
N2 - Kwakhurin (Kwa) is a plant secondary metabolite solely present in Pueraria candollei var. mirifica (P. candollei), which has long been used as a Thai traditional herb for estrogen replacement therapy. Recently, health hazards have arisen in Japan regarding P. candollei-derived products containing potent estrogenic compounds. Therefore, the development of standardization methods for P. candollei materials is an urgent problem requiring resolution. The enzyme-linked immunosorbent assay (ELISA) is an effective analytical technique because it enables the development of sensitive and specific assays of the target compound through antigen−antibody reaction. Here, we produced a monoclonal antibody against Kwa (MAb 11F) by immunizing Kwa-bovine serum albumin (BSA) conjugates prepared using an N,N′-carbonyldiimidazole (CDI) mediated method. Stability and cross-reactivity tests of MAb 11F revealed that the MAb 11F is stable for at least 4 months at 4 °C and is highly specific to Kwa. The detectable concentration range of an indirect competitive ELISA (icELISA) using MAb 11F exhibited values of 1.53–48.8 ng/mL with the limit of detection (LOD) of 1.13 ng/mL. Validation analyses revealed that the developed icELISA is precise, accurate, and reliable enough to be applied to P. candollei-derived samples and products for their standardization.
AB - Kwakhurin (Kwa) is a plant secondary metabolite solely present in Pueraria candollei var. mirifica (P. candollei), which has long been used as a Thai traditional herb for estrogen replacement therapy. Recently, health hazards have arisen in Japan regarding P. candollei-derived products containing potent estrogenic compounds. Therefore, the development of standardization methods for P. candollei materials is an urgent problem requiring resolution. The enzyme-linked immunosorbent assay (ELISA) is an effective analytical technique because it enables the development of sensitive and specific assays of the target compound through antigen−antibody reaction. Here, we produced a monoclonal antibody against Kwa (MAb 11F) by immunizing Kwa-bovine serum albumin (BSA) conjugates prepared using an N,N′-carbonyldiimidazole (CDI) mediated method. Stability and cross-reactivity tests of MAb 11F revealed that the MAb 11F is stable for at least 4 months at 4 °C and is highly specific to Kwa. The detectable concentration range of an indirect competitive ELISA (icELISA) using MAb 11F exhibited values of 1.53–48.8 ng/mL with the limit of detection (LOD) of 1.13 ng/mL. Validation analyses revealed that the developed icELISA is precise, accurate, and reliable enough to be applied to P. candollei-derived samples and products for their standardization.
UR - http://www.scopus.com/inward/record.url?scp=85058674994&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85058674994&partnerID=8YFLogxK
U2 - 10.1016/j.fitote.2018.12.010
DO - 10.1016/j.fitote.2018.12.010
M3 - Article
VL - 133
SP - 23
EP - 28
JO - Fitoterapia
JF - Fitoterapia
SN - 0367-326X
ER -